免疫缺陷、着丝粒不稳定和面部异常综合征家系永生化淋巴细胞系的建立

Establishment of immortalized lymphocyte lines in an immunodeficiency,centromeric instability and facial anomalies syndrome family

目的探讨建立免疫缺陷、着丝粒不稳定和面部异常(ICF)综合征家系永生化淋巴细胞系的方法,为保存这类罕见病家系完整的基因组进行长期研究提供材料。方法应用EB病毒转染ICF综合征家系外周血淋巴细胞,光学显微镜下观察B95-8细胞系及永生化淋巴细胞系形态特征,流式细胞仪检测ICF综合征家系B细胞表达水平,用台盼蓝检测冻存后复苏细胞的活性。结果ICF综合征家系中患儿存在B细胞活化障碍,选择最佳浓度细胞进行EB病毒转染,成功获得永生化淋巴细胞系。永生化淋巴细胞系冻存后复苏,增殖活跃,可继续传代培养,细胞活性大于90%。结论ICF综合征家系体外永生化淋巴细胞系方法的建立,为分子水平上揭示罕见病家系的发病机制提供材料。

Objective To establish immortalized lymphocyte lines and preserve genome in an immunodeficiency,centromeric instability and facial(ICF)abnormalies syndrome family.Methods Immortalized lymphocyte lines were established by EB virus transformation of lymphocyte.The morphological characteristics of B95-8 cell lines were observed under microscope.Flow cytometry was used to detect the expression level of B cells.Trypan blue was used to detect the activity of recovered cells.Results ICF syndrome have B cell activation disorder with children.The best concentration of cells were selected for EB virus transfection.They continued to grow after passaging in the culture and more than 90%cells remain alive after thawing.Conclusion This study provides a method for establishing immortalized lymphocyte lines in vitro from an ICF syndrome family.